UA Biophysics:Protocols:Kanamycin

Kanamycin disulfate salt 25 mg/ml

Kanamycin acts by binding to the 70S ribosomal subunit, inhibiting translocation and eliciting miscoding. Is effective against gram-negative and gram-positive bacteria, and mycoplasma.

Materials

• Kanamycin Disulfate Salt From Streptom K1876-5G
• Sterilized ddH₂O
• Syringe filter
• Syringe 50 ml

Protocol

1. According to desired amount of antibiotics, calculate the amount in grams that has to be weighed. For example, for 10mL of 25mg/mL of Ampicillin: 25mg/mL*10mL=250mg
2. Kanamycin is soluble in ddH₂O.
3. Filter the solution using a 0.2 µm syringe filter.
4. Divide in aliquots of 500μL
5. Store at -20 °C. up to 4-6 months

It is best to avoid repeated freeze/thaw cycles of the kanamycin aliquots. Kanamycin is light sensitive. Keep tightly closed in light-resistant containers. "Solutions are stable at 37°C for approximately 5 days. Aqueous stock solutions can be stored at 2-8°C for long term storage", Sigma. Plates with LB can be stored one month.

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